Abstract

1425

Random periareolar fine needle aspiration (RPFNA) and ductal lavage (DL) are two techniques for procurement of breast epithelium for both risk assessment and measurement of response endpoints in prevention trials. An important objective of an NCI sponsored multi-institutional phase II prevention trial was to compare the efficacy of the two techniques for acquiring tissue sufficient for biomarker studies; as well as comparison of the results of the biomarker studies. Premenopausal women at high risk for development of breast cancer, with a normal breast exam and mammogram during the follicular portion of the menstrual cycle, underwent a NAF attempt and if successful, a DL attempt of NAF producing ducts. Effluent from all lavaged ducts was pooled in Cytolyt™ with 1% formalin. After a 30-minute rest, RPFNA was performed and material from both breasts pooled in formalin modified Cytolyt™. RPFNA was performed regardless of whether NAF was produced. Four slides (ThinPrep) were made from both the DL and RPFNA specimens for cytomorphology and for immunocytochemistry (ICC) of Ki-67, COX-2, and ER. For Ki-67 assessment, a minimum of 500 cells on the slide was required; for COX-2 and ER, a minimum of 100 cells was required for each. One hundred ten subjects (median age =40) were enrolled in the study. Of the 110 RPFNA attempts, all produced adequate cells for cytomorphologic evaluation, but only 90 (82%), 91 (83%), and 103 (94%) had adequate cells for ICC assessment of Ki-67, COX-2, and ER, respectively. Seventy-six subjects produced NAF and underwent a DL attempt. Of the 67 successful DL attempts (specimen obtained), 64 produced >10 cells for cytomorphologic evaluation, but only 46 (72%), 46 (72%), and 55 (86%) had adequate cells for Ki-67, COX-2, and ER, respectively. For the 64 subjects with sufficient cells by DL, 25 had identical cytomorphology by the two procedures; 15 had more abnormal findings in the DL specimen, and 24 had more abnormal findings in the RPFNA specimen. Restricting analysis to subjects with assessments available for both procedures, 40, 43, and 53 cases were evaluable for Ki-67, COX-2, and ER, respectively. ICC expression was higher in the RPFNA specimen compared to the DL specimens for Ki-67 (median percent positive cells: 2.8 vs. 0.6, p<0.001, Wilcoxon Signed Ranks Test), COX-2 (median intensity score: 0.8 vs 0.1, p=0.003), and ER (median intensity score: 0.2 vs 0, p=0.001). In summary, when guided by the production of NAF, DL produced fewer evaluable specimens than RPFNA. Moreover, while cytomorphology was comparable, immunocytochemical expression of Ki-67, COX-2, and ER was less in DL specimens compared to RPFNA specimens. Thus, the RPFNA procedure would appear preferable for obtaining cellular specimens for use in breast cancer prevention trials of premenopausal women. Supported by contract N01-CN-15135 from Division of Cancer Prevention, NCI, NIH.